Premium
Development of leptin mutants with enhanced bioefficacy and stability
Author(s) -
Park Hyunju,
Kim Jongsun
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.564.2
Subject(s) - leptin , medicine , endocrinology , leptin receptor , mutant , in vivo , chemistry , biology , obesity , biochemistry , gene , microbiology and biotechnology
Leptin is a protein hormone produced by adipocytes. It is secreted into the blood stream and plays a key role in regulating body energy homeostasis by inhibiting feeding behavior followed by weight loss. Because protein stability and bioefficacy are major problems in the biotechnology and biopharmaceutical industries, if leptin can be used for the therapeutic proteins, the stability and bioefficacy are needed to be improved. In this study, we mutanted the surface residues of leptin in a systematic way by displaying 3D structure of leptin on pymol program. We compared the biological activities of leptin and mutant leptin ex vivo using MCF7 breast cancer cell line and in vivo using a male C57Bl mouse model. Mutant leptin treated MCF7 cells displayed better cell proliferation than wild type leptin. The biological activities of leptin and mutant leptin were also tested in a semi‐quantitative STAT3 and ERK phosphorylation bioassay. A representative experiment showed at least 2‐fold higher activity in mutant leptin treated group. To assess the in vivo activity, each group of mice was treated with saline, leptin, and mutant leptin for 20 days and the differences in body weight, food intake, abdominal fat contents, and TG concentration were measured. Mutant leptin appeared to decrease the body weight and food intake in male C57Bl mice more significantly than wild type leptin. As a consequence of decreased body weight, the mutant leptin treated mouse group showed decreased abdominal fat contents and low triglyceride (TG) concentration. These results indicate that the mutant leptin is more effective than wild type leptin.